User guide

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Fragment Library Preparation Using the AB Library Builder
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System: 5500 Series SOLiD
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Systems User Guide
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APPENDIXD
Library Construction
Oligonucleotide Sequences
PCR Primer and adaptor sequences
Note: The internal adaptor used for DNA fragment libraries is different from the
internal adaptor used for RNA libraries.
Note:The “~” is a phosphorothioate bond, which protects a sequence from nucleases.
Barcoded adaptor sequences
Adaptor and primer sequences Length (nt)
P1-T Adaptor, 50 µM
5′-CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGA~T-3′
5′-TCACCGACTGCCCATAGAGAGGAAAGCGGAGGCGTAGTGG~C~C-3′
41
42
Standard Adaptor, 50 µM
5′-CGCCTTGGCCGTACAGCAG-3′
5′-CTGCCCCGGGTTCCTCATTCTCTGTGTAAGAGGCTGCTGTACGGCCAAGGCGT-3′
19
53
Library PCR Primer 1, 50 µM
5′-CCACTACGCCTCCGCTTTCCTCTCTATG-3′ 28
Library PCR Primer 2, 50 µM
5′-CTGCCCCGGGTTCCTCATTCT-3′ 21
Barcoded adaptor sequence Length (nt)
Barcode-T-001, 50 µM
5′-CGCCTTGGCCGTACAGCAG-3′
5′-CTGCCCCGGGTTCCTCATTCTCTGTGTAAGAGGCTGCTGTACGGCCAAGGCGT-3′
19
53
Barcode-T-002, 50 µM
5′-CGCCTTGGCCGTACAGCAG3′
5′-CTGCCCCGGGTTCCTCATTCTCTAGGGAGTGGTCTGCTGTACGGCCAAGGCGT-3′
19
53
Barcode-T-003, 50 µM
5′-CGCCTTGGCCGTACAGCAG-3′
5′-CTGCCCCGGGTTCCTCATTCTCTATAGGTTATACTGCTGTACGGCCAAGGCGT-3′
19
53